Objective To investigate the application of BPDE-albumin adducts as monitoring biomarkers for coke oven workers exposed to polycyclic aromatic hydrocarbons （PAHs） and to explore possible relationship between BPDE-al...Objective To investigate the application of BPDE-albumin adducts as monitoring biomarkers for coke oven workers exposed to polycyclic aromatic hydrocarbons （PAHs） and to explore possible relationship between BPDE-albumin adducts and urinary 1-hydroxypyrene （1-OHP） levels in them. Methods Thirty-seven coke oven workers from a coke plant and 47 controls without the occupational exposure to PAHs were recruited in this study. The levels of plasma BPDE-albumin adducts and urinary 1-OHP were analyzed using high performance liquid chromatography. Results The median levels of BPDE-albumin adducts （42.10 fmol/mg albumin） and urinary 1-OHP （5.46 μmol/mol creatinine） were significantly higher in coke oven workers than in controls （14.16 fmol/mg albumin, 2.96μmol/mol creatinine, respectively; P〈0.01）. Multiple logistic regression analysis showed that coke oven workers were at higher risk of having BPDE-albumin adduct levels above 25.30 μmol/mg albumin （OR=1.79, P〈0.01） and urinary 1-OHP levels above 4.13 μmol/mol creatinine （OR=2.45, P〈0.05）. There was a positive correlation between the levels of BPDE-albumin adducts and urinary 1-OHP in all subjects （rs=0.349, P〈0.01）. Conclusion BPDE-albumin adduct is a useful biomarker for monitoring long-term exposure to PAHs, and plasma BPDE-albumin adducts level is significantly correlated to urinary 1-OHP levels in coke oven workers.展开更多
The high-molecular weight polycyclic aromatic hydrocarbons(PAHs) pyrene and typical long chain alkane nhexadecane are both difficult to degrade. In this study, n-hexadecane and pyrene degrading strain Rhodococcus sp. ...The high-molecular weight polycyclic aromatic hydrocarbons(PAHs) pyrene and typical long chain alkane nhexadecane are both difficult to degrade. In this study, n-hexadecane and pyrene degrading strain Rhodococcus sp. T1 was isolated from oil contaminated soil. Strain T1 could remove 90.81% n-hexadecane(2 vol%) and 42.79% pyrene(200 mg·L^(-1)) as a single carbon within 5 days, respectively. Comparatively, the degradation of pyrene increased to 60.63%, but the degradation of n-hexadecane decreased to 87.55% when these compounds were mixed. Additionally, identification and analysis of degradation metabolites of Rhodococcus sp. T1 in the above experiments showed that there were significant changes in alanine, methylamine, citric acid and heptadecanoic acid between sole and dual substrate degradation. The optimal conditions for degradation were then determined based on analysis of the pH, salinity, additional nutrient sources and liquid surface activity.Under the optimal conditions of pH 7.0, 35 °C, 0.5% NaCl, 5 mg·L^(-1) of yeast extract and 90 mg·L^(-1) of surfactant,the degradation increased in single or dual carbon sources. To our knowledge, this is the first study to discuss metabolite changes in Rhodococcus sp. T1 using sole substrate and dual substrate to enhance the long-chain alkanes and PAHs degradation potential.展开更多
Objective To investigate the roles of the cyclin D1/CDK4 and E2F-1/4 pathways and compare their work patterns in cell cycle changes induced by different doses of B[a]E Methods Human embryo lung fibroblasts （HELFs） w...Objective To investigate the roles of the cyclin D1/CDK4 and E2F-1/4 pathways and compare their work patterns in cell cycle changes induced by different doses of B[a]E Methods Human embryo lung fibroblasts （HELFs） were treated with 2 μmol/L or 100 μmol/L B[a]P which were provided with some characteristics of transformed cells （T-HELFs）. Cyclin D l, CDK4 and E2F-1/4 expressions were determined by Western blotting. Flow cytometry was used to detect the distribution of cell cycle. Results After B[a]P treatment, the proportion of the first gap （G 1） phase cells decreased. CDK4 and E2F-4 expression did not change significantly. In 2 μmol/L treated cells, a marked overexpression of cyclin D1 and E2F-1 was observed. However, in T-HELFs overexpression was limited to cyclin D1 only, and no overexpression of E2F-1 was observed. The decreases of G1 phase in response to B[a]P treatment were blocked in antisense cyclin D1 and antisense CDK4 transfected HELFs （A-D1 and A-K4） and T-HELFs （T-A-D1 and T-A-K4）. After 2 μmol/L B[a]P treatment, overexpression of E2F-1 was attenuated in A-D1, and E2F-4 expression was decreased significantly in A-K4. In T-A-D1 and T-A-K4, E2F-4 expression was increased significantly, compared with T-HELFs. The E2F-1 expression remained unchanged in T-A-D1 and T-A-K4. Condusions Cyclin DI/CDK4-E2F-1/4 pathways work in different patterns in response to low dose and high dose B[a]P treatment. In HELFs treated with 2 μmol/L B[a]P, cyclin D1 positively regulates the E2F-1 expression while CDK4 negatively regulates the E2F-4 expression; however, in HELFs treated with 100 μmol/L B[a]P, both cyclin D1 and CDK4 negatively regulate the E2F-4 expression.展开更多
With high performance liquid chromatograph and fluorospectrometry, 1-hydroxypyrene is identified as a metabolite of pyrene in the urine of dogs after intramuscular injection of pyrene. When three successive doses of 1...With high performance liquid chromatograph and fluorospectrometry, 1-hydroxypyrene is identified as a metabolite of pyrene in the urine of dogs after intramuscular injection of pyrene. When three successive doses of 1,2 and 5 μmol/kg wt. pyrene administered, urinary excretion of 1-hydroxypyrene reaches the peak level between 24th and 48th hours. Excreted 1-hydroxypyrene consists of only about 0.05% of injected pyrene.展开更多
Objective To study the benzo(a)pyrene(B[a]P)-induced mRNA expression of aromatic hydrocarbon receptor(AHR)and cytochrome P4501A1(CYP1A1)genes in rat liver.Methods Rats were injected intraperitoneally with 5,10 and 15m...Objective To study the benzo(a)pyrene(B[a]P)-induced mRNA expression of aromatic hydrocarbon receptor(AHR)and cytochrome P4501A1(CYP1A1)genes in rat liver.Methods Rats were injected intraperitoneally with 5,10 and 15mg/kg of B[a]P.The total RNAs were extracted from rat livers by RNA purification kit,and the mRNA expression of AHR and CYP1A1 genes was determined by reverse transcription polymerase chain reaction(RT-PCR).β-actin was used as the internal control.The mRNA expression of both AHR and CYP1A1 genes was measured at indicated time points(24,48 and 72h)after B[a]P treatment at three different concentrations(5,10 and 15mg/kg).Results The mRNA expression of AHR gene increased in a time-dependent manner at the concentration of 10mg/kg but not at 5 and 15mg/kg of B[a]P.The mRNA expression of CYP1A1 gene differed significantly at 48h and 24h in rat livers treated with 10 and 15mg/kg dosage of B[a]P.The mRNA expression of AHR and CYP1A1 genes increased with B[a]P treatment in a concentration-dependent manner.The time-dependent increase in mRNA expression was shown by AHR but not by CYP1A1 gene with B[a]P(10mg/kg)treatment.Conclusion This study demonstrates that toxic B[a]P increases the mRNA expression of both AHR and CYP1A1 genes in vivo,suggesting that B[a]P may play a role in cancer genesis by this way.展开更多
Objective The present study aimed to test whether exposure to benzo（a）pyrene [B（a）P] affects spatial learning and short-term memory by modulating the expression of the Gria1 and Grin2a glutamate receptor subunit g...Objective The present study aimed to test whether exposure to benzo（a）pyrene [B（a）P] affects spatial learning and short-term memory by modulating the expression of the Gria1 and Grin2a glutamate receptor subunit genes in the hippocampus.Methods Thirty-six 21-24-day-old,male rats were randomly assigned into high-,medium-,and low-dose toxin exposure groups （6.25,2.5,and 1 mg/kg,respectively） and a control group,each containing nine rats.The behavioral performance of adult rats exposed to sub-chronic administration of B（a）P was monitored by learning and memory tests （Morris water maze）.Real-time PCR assays were used to quantify Gria1 and Grin2a gene expression in the hippocampus.Results At medium and high doses,B（a）P impaired spatial learning performance.The crossing-platform-location frequency and the time spent swimming in the platform area,which both relate to short-term memory,were significantly decreased in B（a）P-treated rats compared with controls.The level of Gria1 mRNA increased 2.6-5.9-fold,and the level of Grin2a mRNA increased 10-14.5-fold,with a greater fold increase associated with higher doses of B（a）P.Conclusion We demonstrated that sub-chronic administration of B（a）P inhibits spatial learning and short-term memory,and increases Gria1 and Grin2a expression in the hippocampus.This suggests a relationship of B（a）P exposure levels with Gria1 and Grin2a expression and impairment of short-term and spatial memory.展开更多
Background: Diabetes mellitus is one of the major health problems for people all over the world today. According to international diabetes federation reports, diabetes affects 382 million people worldwide. Environment...Background: Diabetes mellitus is one of the major health problems for people all over the world today. According to international diabetes federation reports, diabetes affects 382 million people worldwide. Environmental pollutants have deleterious effects on glucose metabolism and cause insulin resistance. We aimed to investigate the effects of the environmental pollutants benzo(a)pyrene, and the therapeutic potential of resveratrol. Methods: 20 μM of benzo(a)pyrene was administered after 48 h of resveratrol (80 μM) application for 24 h in INS-1 (832/13) insulinoma cells. The cells were treated with 20 μM benzo(a)pyrene for 24 hours after 48 hours initial preconditions with 10 μM resveratrol. Oxidative stress status, insulin secretion and apoptosis were analyzed by molecular techniques. Results: Though resveratrol increased the antioxidant status which was decreased by benzo(a)pyrene, interestingly, it increased the oxidative status. Resveratrol increased benzo(a)pyrene-depleted reduced glutathione levels to the control level. The mRNA expression levels of beta-cell functions associated with genes insulin-1, insulin-2 and sirtuin-1 were upregulated by resveratrol. Resveratrol treatment elevated the insulin concentration of culture medium, and the mRNA expression of forkhead box protein-1 gene. Resveratrol upregulated benzo(a)pyrene-downregulated p53 gene expression. On the other hand, benzo(a)pyrene-downregulated mRNA expression of B-cell lymphoma-2 was induced by resveratrol treatment. Conclusion: The data showed that resveratrol could reverse the oxidative alterations, functional impairments and the carcinogenetic effects of benzo(a)pyrene in pancreas beta-cells.展开更多
Polycyclic aromatic hydrocarbons (PAHs) induce cytochrome P-450 monoxygenase enzymes that catalyze the formation of DNA adducts. We investigated the effects benzo(α)pyrene (B[α]P) alone or in combination with ethano...Polycyclic aromatic hydrocarbons (PAHs) induce cytochrome P-450 monoxygenase enzymes that catalyze the formation of DNA adducts. We investigated the effects benzo(α)pyrene (B[α]P) alone or in combination with ethanol on normal human keratinocyte (NHK) growth, induction of cytochrome P-4501A1 (CYP1A1), and modulation of these treatments by retinoic acid (RA) in a serum-free culture medium. Growth-arrested confluent NHK serum-free cultures were treated with B[α]P alone or in combination with ethanol and RA. The effects on CYP1A1 enzyme activity were investigated. B[α]P treatment alone was not toxic to post-confluent cells;sub-toxic ethanol stimulated cell growth regardless B[α]P treatment. No CYP1A1 activity was detected in control or ethanol-treated NHK cell cultures. B[α]P alone induced CYP1A1 activity, and B[α]P plus ethanol treatment further enhanced B[α]P-induced CYP1A1 activity. Pretreatment with all-trans-RA (t-RA) abolished ethanol enhancement of CYP1A1 activity. There is a synergistic action of ethanol in combination with PAH on induction of P-450 cytochrome enzymes. By contrast, RA reverses ethanol enhancement implying a role for retinoid therapy in counteracting the risk posed by combined alcohol and PAH exposure on epidermal cell carcinogenesis.展开更多
基金This study was supported by National Key Basic Research and Development Program of China (No.2002CB512905)National Natural Science Foundation of China (No.30371204).
文摘Objective To investigate the application of BPDE-albumin adducts as monitoring biomarkers for coke oven workers exposed to polycyclic aromatic hydrocarbons （PAHs） and to explore possible relationship between BPDE-albumin adducts and urinary 1-hydroxypyrene （1-OHP） levels in them. Methods Thirty-seven coke oven workers from a coke plant and 47 controls without the occupational exposure to PAHs were recruited in this study. The levels of plasma BPDE-albumin adducts and urinary 1-OHP were analyzed using high performance liquid chromatography. Results The median levels of BPDE-albumin adducts （42.10 fmol/mg albumin） and urinary 1-OHP （5.46 μmol/mol creatinine） were significantly higher in coke oven workers than in controls （14.16 fmol/mg albumin, 2.96μmol/mol creatinine, respectively; P〈0.01）. Multiple logistic regression analysis showed that coke oven workers were at higher risk of having BPDE-albumin adduct levels above 25.30 μmol/mg albumin （OR=1.79, P〈0.01） and urinary 1-OHP levels above 4.13 μmol/mol creatinine （OR=2.45, P〈0.05）. There was a positive correlation between the levels of BPDE-albumin adducts and urinary 1-OHP in all subjects （rs=0.349, P〈0.01）. Conclusion BPDE-albumin adduct is a useful biomarker for monitoring long-term exposure to PAHs, and plasma BPDE-albumin adducts level is significantly correlated to urinary 1-OHP levels in coke oven workers.
基金Supported by the National Basic Research Program of China("973" Program:2014CB745100)the National Natural Science Foundation of China(21576197)Tianjin Key Research&Development Program(16YFXTSF00460)
文摘The high-molecular weight polycyclic aromatic hydrocarbons(PAHs) pyrene and typical long chain alkane nhexadecane are both difficult to degrade. In this study, n-hexadecane and pyrene degrading strain Rhodococcus sp. T1 was isolated from oil contaminated soil. Strain T1 could remove 90.81% n-hexadecane(2 vol%) and 42.79% pyrene(200 mg·L^(-1)) as a single carbon within 5 days, respectively. Comparatively, the degradation of pyrene increased to 60.63%, but the degradation of n-hexadecane decreased to 87.55% when these compounds were mixed. Additionally, identification and analysis of degradation metabolites of Rhodococcus sp. T1 in the above experiments showed that there were significant changes in alanine, methylamine, citric acid and heptadecanoic acid between sole and dual substrate degradation. The optimal conditions for degradation were then determined based on analysis of the pH, salinity, additional nutrient sources and liquid surface activity.Under the optimal conditions of pH 7.0, 35 °C, 0.5% NaCl, 5 mg·L^(-1) of yeast extract and 90 mg·L^(-1) of surfactant,the degradation increased in single or dual carbon sources. To our knowledge, this is the first study to discuss metabolite changes in Rhodococcus sp. T1 using sole substrate and dual substrate to enhance the long-chain alkanes and PAHs degradation potential.
基金Grants of National Natural Science Foundation of China (30371206, 30028019)973 National Key Basic Research and Development Program (2002 CB 512905)
文摘Objective To investigate the roles of the cyclin D1/CDK4 and E2F-1/4 pathways and compare their work patterns in cell cycle changes induced by different doses of B[a]E Methods Human embryo lung fibroblasts （HELFs） were treated with 2 μmol/L or 100 μmol/L B[a]P which were provided with some characteristics of transformed cells （T-HELFs）. Cyclin D l, CDK4 and E2F-1/4 expressions were determined by Western blotting. Flow cytometry was used to detect the distribution of cell cycle. Results After B[a]P treatment, the proportion of the first gap （G 1） phase cells decreased. CDK4 and E2F-4 expression did not change significantly. In 2 μmol/L treated cells, a marked overexpression of cyclin D1 and E2F-1 was observed. However, in T-HELFs overexpression was limited to cyclin D1 only, and no overexpression of E2F-1 was observed. The decreases of G1 phase in response to B[a]P treatment were blocked in antisense cyclin D1 and antisense CDK4 transfected HELFs （A-D1 and A-K4） and T-HELFs （T-A-D1 and T-A-K4）. After 2 μmol/L B[a]P treatment, overexpression of E2F-1 was attenuated in A-D1, and E2F-4 expression was decreased significantly in A-K4. In T-A-D1 and T-A-K4, E2F-4 expression was increased significantly, compared with T-HELFs. The E2F-1 expression remained unchanged in T-A-D1 and T-A-K4. Condusions Cyclin DI/CDK4-E2F-1/4 pathways work in different patterns in response to low dose and high dose B[a]P treatment. In HELFs treated with 2 μmol/L B[a]P, cyclin D1 positively regulates the E2F-1 expression while CDK4 negatively regulates the E2F-4 expression; however, in HELFs treated with 100 μmol/L B[a]P, both cyclin D1 and CDK4 negatively regulate the E2F-4 expression.
基金This project is funded by the National Natural Sciences Foundation of China, and partially supported by Beijing Municiple Environmental Protection Bureau.
文摘With high performance liquid chromatograph and fluorospectrometry, 1-hydroxypyrene is identified as a metabolite of pyrene in the urine of dogs after intramuscular injection of pyrene. When three successive doses of 1,2 and 5 μmol/kg wt. pyrene administered, urinary excretion of 1-hydroxypyrene reaches the peak level between 24th and 48th hours. Excreted 1-hydroxypyrene consists of only about 0.05% of injected pyrene.
基金supported by the National Natural Science Foundation of China(No.30660214)the Natural Science Foundation of Inner Mongolia Autonomous Region(No.200607010907)Doctor Initializing Fund Application of Inner Mongolia Medical College(No.2005BQ001)
文摘Objective To study the benzo(a)pyrene(B[a]P)-induced mRNA expression of aromatic hydrocarbon receptor(AHR)and cytochrome P4501A1(CYP1A1)genes in rat liver.Methods Rats were injected intraperitoneally with 5,10 and 15mg/kg of B[a]P.The total RNAs were extracted from rat livers by RNA purification kit,and the mRNA expression of AHR and CYP1A1 genes was determined by reverse transcription polymerase chain reaction(RT-PCR).β-actin was used as the internal control.The mRNA expression of both AHR and CYP1A1 genes was measured at indicated time points(24,48 and 72h)after B[a]P treatment at three different concentrations(5,10 and 15mg/kg).Results The mRNA expression of AHR gene increased in a time-dependent manner at the concentration of 10mg/kg but not at 5 and 15mg/kg of B[a]P.The mRNA expression of CYP1A1 gene differed significantly at 48h and 24h in rat livers treated with 10 and 15mg/kg dosage of B[a]P.The mRNA expression of AHR and CYP1A1 genes increased with B[a]P treatment in a concentration-dependent manner.The time-dependent increase in mRNA expression was shown by AHR but not by CYP1A1 gene with B[a]P(10mg/kg)treatment.Conclusion This study demonstrates that toxic B[a]P increases the mRNA expression of both AHR and CYP1A1 genes in vivo,suggesting that B[a]P may play a role in cancer genesis by this way.
基金support by the National Science Foundation of China (NO.30671744)
文摘Objective The present study aimed to test whether exposure to benzo（a）pyrene [B（a）P] affects spatial learning and short-term memory by modulating the expression of the Gria1 and Grin2a glutamate receptor subunit genes in the hippocampus.Methods Thirty-six 21-24-day-old,male rats were randomly assigned into high-,medium-,and low-dose toxin exposure groups （6.25,2.5,and 1 mg/kg,respectively） and a control group,each containing nine rats.The behavioral performance of adult rats exposed to sub-chronic administration of B（a）P was monitored by learning and memory tests （Morris water maze）.Real-time PCR assays were used to quantify Gria1 and Grin2a gene expression in the hippocampus.Results At medium and high doses,B（a）P impaired spatial learning performance.The crossing-platform-location frequency and the time spent swimming in the platform area,which both relate to short-term memory,were significantly decreased in B（a）P-treated rats compared with controls.The level of Gria1 mRNA increased 2.6-5.9-fold,and the level of Grin2a mRNA increased 10-14.5-fold,with a greater fold increase associated with higher doses of B（a）P.Conclusion We demonstrated that sub-chronic administration of B（a）P inhibits spatial learning and short-term memory,and increases Gria1 and Grin2a expression in the hippocampus.This suggests a relationship of B（a）P exposure levels with Gria1 and Grin2a expression and impairment of short-term and spatial memory.
基金supported by the Scientific Research Projects Unit of Afyon Kocatepe University(Project No:10.TIP.18/MSc Thesis in Medical Biochemistry)NATO-SPS Program(Project No:MYP-G5266).
文摘Background: Diabetes mellitus is one of the major health problems for people all over the world today. According to international diabetes federation reports, diabetes affects 382 million people worldwide. Environmental pollutants have deleterious effects on glucose metabolism and cause insulin resistance. We aimed to investigate the effects of the environmental pollutants benzo(a)pyrene, and the therapeutic potential of resveratrol. Methods: 20 μM of benzo(a)pyrene was administered after 48 h of resveratrol (80 μM) application for 24 h in INS-1 (832/13) insulinoma cells. The cells were treated with 20 μM benzo(a)pyrene for 24 hours after 48 hours initial preconditions with 10 μM resveratrol. Oxidative stress status, insulin secretion and apoptosis were analyzed by molecular techniques. Results: Though resveratrol increased the antioxidant status which was decreased by benzo(a)pyrene, interestingly, it increased the oxidative status. Resveratrol increased benzo(a)pyrene-depleted reduced glutathione levels to the control level. The mRNA expression levels of beta-cell functions associated with genes insulin-1, insulin-2 and sirtuin-1 were upregulated by resveratrol. Resveratrol treatment elevated the insulin concentration of culture medium, and the mRNA expression of forkhead box protein-1 gene. Resveratrol upregulated benzo(a)pyrene-downregulated p53 gene expression. On the other hand, benzo(a)pyrene-downregulated mRNA expression of B-cell lymphoma-2 was induced by resveratrol treatment. Conclusion: The data showed that resveratrol could reverse the oxidative alterations, functional impairments and the carcinogenetic effects of benzo(a)pyrene in pancreas beta-cells.
文摘Polycyclic aromatic hydrocarbons (PAHs) induce cytochrome P-450 monoxygenase enzymes that catalyze the formation of DNA adducts. We investigated the effects benzo(α)pyrene (B[α]P) alone or in combination with ethanol on normal human keratinocyte (NHK) growth, induction of cytochrome P-4501A1 (CYP1A1), and modulation of these treatments by retinoic acid (RA) in a serum-free culture medium. Growth-arrested confluent NHK serum-free cultures were treated with B[α]P alone or in combination with ethanol and RA. The effects on CYP1A1 enzyme activity were investigated. B[α]P treatment alone was not toxic to post-confluent cells;sub-toxic ethanol stimulated cell growth regardless B[α]P treatment. No CYP1A1 activity was detected in control or ethanol-treated NHK cell cultures. B[α]P alone induced CYP1A1 activity, and B[α]P plus ethanol treatment further enhanced B[α]P-induced CYP1A1 activity. Pretreatment with all-trans-RA (t-RA) abolished ethanol enhancement of CYP1A1 activity. There is a synergistic action of ethanol in combination with PAH on induction of P-450 cytochrome enzymes. By contrast, RA reverses ethanol enhancement implying a role for retinoid therapy in counteracting the risk posed by combined alcohol and PAH exposure on epidermal cell carcinogenesis.
